LOCUS       sequence-69022-e        3323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3323)
  AUTHORS   Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S
  TITLE     A modular cloning system for standardized assembly of multigene 
            constructs.
  JOURNAL   PLoS One. 2011 Feb 18;6(2):e16765. doi: 
            10.1371/journal.pone.0016765.
  PUBMED    21364738
REFERENCE   2  (bases 1 to 3323)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2011-02-18"; volume: "6(2):e16765. doi"; pages: " 
            10.1371/journal.pone.0016765"
COMMENT     Sequence Label: pICH50866
FEATURES             Location/Qualifiers
     source          1..3323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..37)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(20..36)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(29..51)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(245..264)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(424..442)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        510..614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             834..1625
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1796..2384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2285..2304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2538..2555
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2672..2693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2708..2713,2714..2731,2732..2738)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2746..2762
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2751..2773
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2770..2786
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2770..2786
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
       61 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
      121 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
      181 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
      241 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
      301 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgacca
      361 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagtcgaa
      421 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
      481 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      601 gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg
      661 gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta
      721 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
      781 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac atcatgaggg
      841 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
      901 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
      961 agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
     1021 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc
     1081 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
     1141 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg
     1201 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
     1261 ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat
     1321 ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg
     1381 agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg
     1441 cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg
     1501 tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg
     1561 cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca
     1621 aataactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1681 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1741 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1801 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1861 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1921 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1981 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2041 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2101 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2161 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2221 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2281 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2341 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2401 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2461 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2521 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     2581 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     2641 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     2701 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     2761 aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg
     2821 actctagagg atccccttga agacaatgcc gaattcgata tcgcacatgt gaccgaggga
     2881 ttgtcttcgt ggaaacttgg gacttcagat tccacccctt gcaagcgtgg aaagccttcc
     2941 gaccaaggga agtgtcggat gtagaggaaa tggaaagttt gttctcagat ggggacttgc
     3001 ttgattgctt cacaagaatg ccagcttatg cggtaaacgc agaggaagat ttagctgcaa
     3061 tcaggaaaac gcccgagatg gatgtcggtc aagaagttaa agaacctgca ggagacagaa
     3121 atcaatactc aaaccctgca gaaactttcc tcaacaagct ccacaggaaa cacagtaggg
     3181 aggtgaaaca ccaggccgca aagaaagcta aacgcctagc tgaaatccag gagtcaatga
     3241 gagctgaagg tgatgccgaa ccaaatgaaa taagcggggg catgggggca atacccagca
     3301 acgccgaact tcctgggccc aaa
//
